LOCUS       sequence-429709-        5998 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses motor domain of KIF17 fused to FIREmate in mamallian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5998)
  AUTHORS   Bottone S, Joliot O, Cakil ZV, El Hajji L, Rakotoarison LM, 
            Boncompain G, Perez F, Gautier A
  TITLE     A fluorogenic chemically induced dimerization technology for 
            controlling, imaging and sensing protein proximity.
  JOURNAL   Nat Methods. 2023 Aug 28. doi: 10.1038/s41592-023-01988-8.
  PUBMED    37640938
REFERENCE   2  (bases 1 to 5998)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Methods. 2023 Aug 28. doi: 10.1038/s41592-023-01988-8."
COMMENT     Sequence Label: FP5369 KIF17MD-flag- FIREmate
FEATURES             Location/Qualifiers
     source          1..5998
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        65..368
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        369..572
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     523..543
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2292..2315
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2790..2911
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2827..2846)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2881..2900
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2918..3373)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3055..3076)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3267..3286
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3400..3504
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3506..3863
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3506..3697
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3529..3549)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3714..3849
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3898..4692
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3898..4692
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3952..3971)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4562..4581
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4880..4899)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4924..4971
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5300..5888
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5789..5808
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcattagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
       61 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
      121 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
      181 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
      241 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
      301 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
      361 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
      421 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
      481 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
      541 gtgtacggtg ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc
      601 gctaccggac tcagatctcg agctcaagct tcgaattcat ggcctccgag gcggtgaagg
      661 ttgtcgtgcg ctgccgtccc atgaaccagc gggagcgaga gctgcgctgc cagcccgtgg
      721 tgactgtgga ctgcgcgcgc gcccagtgct gcatccagaa cccgggcgcc gccgacgagc
      781 cgcccaagca gttcaccttc gacggcgcct accacgtgga ccacgtcacc gagcagatct
      841 acaacgagat cgcctatccg ctggtggagg gcgtcactga gggctacaat ggcaccatct
      901 ttgcctacgg ccagacaggc agcgggaagt ccttcaccat gcagggcctg ccggatccgc
      961 cctcccagag aggcatcatc cccagggcct tcgagcacgt gttcgagagc gtccagtgtg
     1021 cagagaacac taagttcctg gtccgggcct cctacctgga gatctacaat gaagatgtcc
     1081 gggacctcct tggggctgac accaagcaga agctggagct gaaggagcac ccagagaagg
     1141 gcgtgtacgt gaaggggctg tccatgcaca cggtgcacag cgtggcccag tgtgagcaca
     1201 tcatggagac tggctggaag aaccgttcgg tcggctacac gctgatgaac aaggattcct
     1261 cacgctcgca ctccatcttc accatcagca tcgagatgtc tgccgtggat gagcggggca
     1321 aggaccacct ccgggcgggc aagctgaacc tggtggacct ggcgggcagc gagcggcagt
     1381 ccaagaccgg ggccacgggc gagcggctca aggaggccac caagatcaac ctgtcgctct
     1441 cggcactggg caatgtcatc tcggcgctgg tggacgggcg ctgtaagcac gtcccctacc
     1501 gtgactcgaa gctgacgcgg ctgctgcagg actcactggg cggcaacacc aagacgctca
     1561 tggtggcctg cctgtcgcct gcggacaaca actacgatga gacactcagc acgctgcgct
     1621 acgccaaccg ggccaagaac atcaggaaca agccgcgcat caatgaggac cccaaggatg
     1681 cgctgcttcg cgagtaccag gaggagatca agaagctcaa ggccatcctg acacagcaga
     1741 tgagccccag cagcctgtca gccctgctgt ccaggcaggt gcccccagac cctgtgcagg
     1801 tggaggagaa gctgttgccc caacctgtga tccagcatga catggaggcc gagaagcagc
     1861 tgatccggga ggagtatgaa gagcgcctgg cccggctgaa agccgactat aaggccgagc
     1921 aggagtctcg ggccaggctg gaggaagaca tcactgccat gcgcaactca tatgacgtca
     1981 ggctgtccac gctggaggag aacctgcgga aggagacaga ggctgtcctg caggtgggag
     2041 tcctctacaa ggctgaggtc atgtccaggg ctgagtttgc cagcagcgct gagtacccgc
     2101 ctgcttttca gtatgagaca gtggtgaaac ccaaggtctt ctccacgact gacactctgc
     2161 ccagtgacga tgtctccaag actcaggttt cctccaggtt tgcggagctg cccaaggtgg
     2221 aaccctccaa atctgagatt tctctgggct ccagtgagtc atcctcgctc gaagaaaccc
     2281 ttgcggccgc tgattacaag gatgacgatg acaagttaat taaagagcat gttgcctttg
     2341 gcagtgagga catcgagaac actctggcca atatggacga cgaacaactg gataggttgg
     2401 cctttggcgt aattcagctc gatggtgacg ggaatatcct gctgtacaat gctgctgaag
     2461 gggacatcac tggcagagat cccaaacagg tgattgggaa gaacttcttc aaggatgttg
     2521 cacctggaac ggatactccc gagttttacg gcaaattcaa ggaaggcgca gcgtcaggga
     2581 atctgaacac catgttcgaa tggacgatac cgacaagcag gggaccaacc aaggtcaagg
     2641 tgcacttgaa gaaagccctt tcctgaggcc ggccgcgact ctagatcata atcagccata
     2701 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     2761 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     2821 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     2881 gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg
     2941 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     3001 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     3061 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     3121 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     3181 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     3241 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     3301 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     3361 ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     3421 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     3481 caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc
     3541 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3601 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3661 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     3721 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     3781 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     3841 ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg
     3901 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     3961 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     4021 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa
     4081 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     4141 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     4201 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     4261 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     4321 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     4381 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc
     4441 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     4501 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     4561 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     4621 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     4681 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     4741 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     4801 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     4861 accctagggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta
     4921 tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg
     4981 gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat
     5041 acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc
     5101 tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt
     5161 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     5221 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     5281 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     5341 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     5401 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     5461 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     5521 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     5581 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     5641 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     5701 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     5761 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     5821 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     5881 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     5941 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccat
//
